Pulsed field separation of large supercoiled and open-circular DNAs and its application to bacterial artificial chromosome cloning

1995 ◽  
Vol 16 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Min Wang ◽  
Eric Lai
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Artificial Chromosome ◽  
Pulsed Field ◽  
Open Circular ◽  
Circular Dnas ◽  
Bacterial Artificial Chromosome Cloning ◽  
Field Separation

scholarly journals Bacterial artificial chromosome cloning and mapping of a 630-kb human extrachromosomal structure.

1996 ◽  
Vol 6 (7) ◽  
pp. 612-619 ◽  
Author(s):  
M Wang ◽  
S Shouse ◽  
B Lipes ◽  
U J Kim ◽  
H Shizuya ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Artificial Chromosome ◽  
Bacterial Artificial Chromosome Cloning

A bacterial artificial chromosome based physical map of the Ustilago maydis genome

Genome ◽  
2005 ◽  
Vol 48 (2) ◽  
pp. 207-216 ◽  
Author(s):  
Khalid Meksem ◽  
Jeffry Shultz ◽  
Faiza Tebbji ◽  
Aziz Jamai ◽  
Jürgen Henrich ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Gel Electrophoresis ◽  
Physical Map ◽  
Artificial Chromosome ◽  
Ustilago Maydis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Artificial Chromosomes ◽  
Bac Clones ◽  
Copy Dna

Ustilago maydis, a basidiomycete, is a model organism among phytopathogenic fungi. A physical map of U. maydis strain 521 was developed from bacterial artificial chromosome (BAC) clones. BAC fingerprints used polyacrylamide gel electrophoresis to separate restriction fragments. Fragments were labeled at the HindIII site and codigested with HaeIII to reduce fragments to 50–750 bp. Contiguous overlapping sets of clones (contigs) were assembled at nine stringencies (from P ≤ 1 x 10–6 to 1 x 10–24). Each assembly nucleated contigs with different percentages of bands overlapping between clones (from 20% to 97%). The number of clones per contig decreased linearly from 41 to 12 from P ≤ 1 x 10–7 to 1 x 10–12. The number of separate contigs increased from 56 to 150 over the same range. A hybridization-based physical map of the same BAC clones was compared with the fingerprint contigs built at P ≤ 1 × 10–7. The two methods provided consistent physical maps that were largely validated by genome sequence. The combined hybridization and fingerprint physical map provided a minimum tile path composed of 258 BAC clones (18–20 Mbp) distributed among 28 merged contigs. The genome of U. maydis was estimated to be 20.5 Mbp by pulsed-field gel electrophoresis and 24 Mbp by BAC fingerprints. There were 23 separate chromosomes inferred by both pulsed-field gel electrophoresis and fingerprint contigs. Only 11 of the tile path BAC clones contained recognizable centromere, telomere, and subtelomere repeats (high-copy DNA), suggesting that repeats caused some false merges. There were 247 tile path BAC clones that encompassed about 17.5 Mbp of low-copy DNA sequence. BAC clones are available for repeat and unique gene cluster analysis including tDNA-mediated transformation. Program FingerPrint Contigs maps aligned with each chromosome can be viewed at http://www.siu.edu/~meksem/ustilago_maydis/.Key words: Ustilago maydis, physical map, bacterial artificial chromosomes, whole-genome sequencing.

Development of a novel bacterial artificial chromosome cloning system for functional studies

Plasmid ◽  
2003 ◽  
Vol 49 (2) ◽  
pp. 184-187 ◽  
Author(s):  
Keith Al-Hasani ◽  
Kim Simpfendorfer ◽  
Hady Wardan ◽  
Jim Vadolas ◽  
Faten Zaibak ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Artificial Chromosome ◽  
Cloning System ◽  
Functional Studies ◽  
Bacterial Artificial Chromosome Cloning

scholarly journals Development of bovine herpesvirus 4 as an expression vector using bacterial artificial chromosome cloning

2005 ◽  
Vol 86 (4) ◽  
pp. 907-917 ◽  
Author(s):  
L. Gillet ◽  
V. Daix ◽  
G. Donofrio ◽  
M. Wagner ◽  
U. H. Koszinowski ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Expression Vector ◽  
Bovine Herpesvirus ◽  
Artificial Chromosome ◽  
Complement Protein ◽  
Bovine Herpesvirus 4 ◽  
Infected Cells ◽  
Bac Cloning ◽  
Herpesvirus 4 ◽  
Bacterial Artificial Chromosome Cloning

Several features make bovine herpesvirus 4 (BoHV-4) attractive as a backbone for use as a viral expression vector and/or as a model to study gammaherpesvirus biology. However, these developments have been impeded by the difficulty in manipulating its large genome using classical homologous recombination in eukaryotic cells. In the present study, the feasibility of exploiting bacterial artificial chromosome (BAC) cloning and prokaryotic recombination technology for production of BoHV-4 recombinants was explored. Firstly, the BoHV-4 genome was BAC cloned using two potential insertion sites. Both sites of insertion gave rise to BoHV-4 BAC clones stably maintained in bacteria and able to regenerate virions when transfected into permissive cells. Reconstituted virus replicated comparably to wild-type parental virus and the loxP-flanked BAC cassette was excised by growing them on permissive cells stably expressing Cre recombinase. Secondly, BoHV-4 recombinants expressing Ixodes ricinus anti-complement protein I or II (IRAC I/II) were produced using a two-step mutagenesis procedure in Escherichia coli. Both recombinants induced expression of high levels of functional IRAC molecules in the supernatant of infected cells. This study demonstrates that BAC cloning and prokaryotic recombination technology are powerful tools for the development of BoHV-4 as an expression vector and for further fundamental studies of this gammaherpesvirus.

scholarly journals Targeting CreERT2 expression to keratin 8-expressing murine simple epithelia using bacterial artificial chromosome transgenesis

2012 ◽  
Vol 21 (5) ◽  
pp. 1117-1123 ◽  
Author(s):  
Li Zhang ◽  
Boyu Zhang ◽  
Sang Jun Han ◽  
Amy N. Shore ◽  
Jeffrey M. Rosen ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Artificial Chromosome ◽  
Keratin 8

scholarly journals Revising the Chromosome-Specific Probes of White Hawk (Leucopternis albicollis)

Proceedings ◽  
2020 ◽  
Vol 76 (1) ◽  
pp. 1
Author(s):  
Ivanete Furo ◽  
Rafael Kretschmer ◽  
Jorge Pereira ◽  
Darren Griffin ◽  
Rebecca O’Connor ◽  
...  
Keyword(s):  
Bacterial Artificial Chromosome ◽  
Artificial Chromosome ◽  
Bird Of Prey ◽  
Inaccurate Data ◽  
Intrachromosomal Rearrangements

Leucopternis albicollis is a diurnal bird of prey with extensive karyotype reorganization. Chromosome-specific probes from this species have been used successfully to detect intrachromosomal rearrangements in different species of bird since 2010. However, some gaps were detected in this first set of probes. Here, we have obtained a new set of whole chromosome probes in order to improve the previous one; also, we have performed experiments using bacterial artificial chromosome (BAC) from chicken microchromosomes. Our results demonstrated that the microchromosomes were involved in fusion events. In addition, a new nomenclature has been proposed for the new set of probes and some inaccurate data were corrected.

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